1887

Abstract

Acyl carrier proteins (ACPs) are required for the transfer of acyl intermediates during fatty acid and polyketide syntheses. In 1021 there are five known ACPs: AcpP, NodF, AcpXL, the ACP domain in RkpA and SMb20651. The genome sequence of 1021 also reveals the ORF SMc01553, annotated as a putative ACP. is part of a 6.6 kb DNA region that is duplicated in the chromosome and in the pSymb plasmid, the result of a recent duplication event. SMc01553 overexpressed in was labelled with [H]-alanine, a biosynthetic building block of the 4′-phosphopantetheine prosthetic group of ACPs. The purified SMc01553 was modified with 4′-phosphopantetheine in the presence of holo-ACP synthase, and this modification resulted in a major conformational change of the protein structure, since the holo-form runs faster in native PAGE than the apo-form. SMc01553 could not be loaded with a malonyl group by malonyl-CoA-ACP transacylase from . Using RT-PCR we could show the presence of mRNA for SMc01553 and of the duplicated ORF SMb22007 in cultures of . However, a mutant in which the two duplicated regions were deleted did not show any different phenotype with respect to the wild-type in the free-living or symbiotic lifestyle.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.033480-0
2010-01-01
2019-11-13
Loading full text...

Full text loading...

/deliver/fulltext/micro/156/1/230.html?itemId=/content/journal/micro/10.1099/mic.0.033480-0&mimeType=html&fmt=ahah

References

  1. Altschul, S. F., Madden, T. L., Schäfer, A. A., Zhang, J., Zhang, Z., Miller, W. & Lipman, D. J. ( 1997; ). Gapped blast and psi-blast: a new generation of protein database search programs. Nucleic Acids Res 25, 3389–3402.[CrossRef]
    [Google Scholar]
  2. Ardourel, M., Demont, N., Debellé, F., Maillet, F., de Billy, F., Promé, J. C., Dénarié, J. & Truchet, G. ( 1994; ). Rhizobium meliloti lipooligosaccharide nodulation factors: different structural requirements for bacterial entry into target root hair cells and induction of plant symbiotic developmental responses. Plant Cell 6, 1357–1374.[CrossRef]
    [Google Scholar]
  3. Barnett, M. J., Toman, C. J., Fisher, R. F. & Long, S. R. ( 2004; ). A dual-genome symbiosis chip for coordinate study of signal exchange and development in a prokaryote–host interaction. Proc Natl Acad Sci U S A 101, 16636–16641.[CrossRef]
    [Google Scholar]
  4. Becker, A., Fraysse, N. & Sharypova, L. ( 2005; ). Recent advances in studies on structure and symbiosis-related function of rhizobial K-antigens and lipopolysaccharides. Mol Plant Microbe Interact 18, 899–905.[CrossRef]
    [Google Scholar]
  5. Beringer, J. E. ( 1974; ). R factor transfer in Rhizobium leguminosarum. J Gen Microbiol 84, 188–198.[CrossRef]
    [Google Scholar]
  6. Brozek, K. A., Carlson, R. W. & Raetz, C. R. ( 1996; ). A special acyl carrier protein for transferring long hydroxylated fatty acids to lipid A in Rhizobium. J Biol Chem 271, 32126–32136.[CrossRef]
    [Google Scholar]
  7. Burris, R. H. ( 1972; ). Nitrogen fixation assay–methods and techniques. Methods Enzymol 24, 415–431.
    [Google Scholar]
  8. Byers, D. M. & Gong, H. ( 2007; ). Acyl carrier protein: structure–function relationships in a conserved multifunctional protein family. Biochem Cell Biol 85, 649–662.[CrossRef]
    [Google Scholar]
  9. Chan, Y. A., Podevels, A. M., Kevany, B. M. & Thomas, M. G. ( 2009; ). Biosynthesis of polyketide synthase extender units. Nat Prod Rep 26, 90–114.[CrossRef]
    [Google Scholar]
  10. Dulley, J. R. & Grieve, P. A. ( 1975; ). A simple technique for eliminating interference by detergents in the Lowry method of protein determination. Anal Biochem 64, 136–141.[CrossRef]
    [Google Scholar]
  11. Epple, G., van der Drift, K. M. G. M., Thomas-Oates, J. & Geiger, O. ( 1998; ). Characterization of a novel acyl carrier protein, RkpF, encoded by an operon involved in capsular polysaccharide biosynthesis in Sinorhizobium meliloti. J Bacteriol 180, 4950–4954.
    [Google Scholar]
  12. Ferguson, G. P., Roop, R. M., II & Walker, G. C. ( 2002; ). Deficiency of Sinorhizobium meliloti bacA mutant in alfalfa symbiosis correlates with the alteration of cell envelope. J Bacteriol 184, 5625–5632.[CrossRef]
    [Google Scholar]
  13. Ferguson, G. P., Datta, A., Carlson, R. W. & Walker, G. C. ( 2005; ). Importance of unusually modified lipid A in Sinorhizobium stress resistance and legume symbiosis. Mol Microbiol 56, 68–80.[CrossRef]
    [Google Scholar]
  14. Florova, G., Kazanina, G. & Reynolds, K. A. ( 2002; ). Enzymes involved in fatty acid and polyketide biosynthesis in Streptomyces glaucescens: role of FabH and FabD and their acyl carrier protein specificity. Biochemistry 41, 10462–10471.[CrossRef]
    [Google Scholar]
  15. Flugel, R. S., Hwangbo, Y., Lambalot, R. H., Cronan, J. E. & Walsh, C. T. ( 2000; ). Holo-(acyl carrier protein) synthase and phosphopantetheinyl transfer in Escherichia coli. J Biol Chem 275, 959–978.[CrossRef]
    [Google Scholar]
  16. Galibert, F., Finan, T. M., Long, S. R., Pühler, A., Abola, P., Ampe, F., Barloy-Hubler, F., Barnett, M. J., Becker, A. & other authors ( 2001; ). The composite genome of the legume symbiont Sinorhizobium meliloti. Science 293, 668–672.[CrossRef]
    [Google Scholar]
  17. Geiger, O. & López-Lara, I. M. ( 2002; ). Rhizobial acyl carrier proteins and their roles in the formation of bacterial cell surface components that are required for the development of nitrogen-fixing root nodules on legume hosts. FEMS Microbiol Lett 208, 153–162.[CrossRef]
    [Google Scholar]
  18. Geiger, O., Spaink, H. P. & Kennedy, E. P. ( 1991; ). Isolation of the Rhizobium leguminosarum NodF nodulation protein: NodF carries a 4′-phosphopantetheine prosthetic group. J Bacteriol 173, 2872–2878.
    [Google Scholar]
  19. Guo, X., Flores, M., Mavingui, P., Fuentes, S. I., Hernández, G., Dávila, G. & Palacios, R. ( 2003; ). Natural genomic design in Sinorhizobium meliloti: novel genomic architectures. Genome Res 13, 1810–1817.
    [Google Scholar]
  20. Haag, A. F., Wehmeier, S., Beck, S., Marlow, V. L., Fletcher, V., James, E. K. & Ferguson, G. P. ( 2009; ). The Sinorhizobium meliloti LpxXL and AcpXL proteins play important roles in bacteroid development within alfalfa. J Bacteriol 191, 4681–4686.[CrossRef]
    [Google Scholar]
  21. Hofmann, K. ( 2000; ). A superfamily of membrane-bound O-acyltransferases with implications for Wnt signaling. Trends Biochem Sci 25, 111–112.[CrossRef]
    [Google Scholar]
  22. Jackowski, S. & Rock, C. O. ( 1983; ). Ratio of active to inactive forms of acyl carrier protein in Escherichia coli. J Biol Chem 258, 15186–15191.
    [Google Scholar]
  23. Lambalot, R. H., Gehring, A. M., Flugel, R. S., Zuber, P., LaCelle, M., Marahiel, M. A., Reid, R., Khosla, C. & Walsh, C. T. ( 1996; ). A new enzyme superfamily: the phosphopantetheinyl transferases. Chem Biol 3, 923–936.[CrossRef]
    [Google Scholar]
  24. López-Lara, I. M. & Geiger, O. ( 2000; ). Expression and purification of four different acyl carrier proteins. Microbiology 146, 839–849.
    [Google Scholar]
  25. Miller, J. H. ( 1972; ). Experiments in Molecular Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  26. Olivares, J., Casadesus, J. & Bedmar, E. J. ( 1980; ). Method for testing degree of infectivity of Rhizobium meliloti strains. Appl Environ Microbiol 39, 967–970.
    [Google Scholar]
  27. Østeras, M., Boncompagni, E., Vincent, N., Poggi, M. C. & Le Rudulier, D. ( 1998; ). Presence of a gene encoding choline sulfatase in Sinorhizobium meliloti bet operon: choline-O-sulfate is metabolised into glycine betaine. Proc Natl Acad Sci U S A 95, 11394–11399.[CrossRef]
    [Google Scholar]
  28. Ramos-Vega, A. L., Dávila-Martínez, Y., Sohlenkamp, C., Contreras-Martínez, S., Encarnación, S., Geiger, O. & López-Lara, I. M. ( 2009; ). SMb20651 is another acyl carrier protein from Sinorhizobium meliloti. Microbiology 155, 257–267.[CrossRef]
    [Google Scholar]
  29. Rawlings, M. & Cronan, J. E. ( 1992; ). The gene encoding Escherichia coli acyl carrier protein lies within a cluster of fatty acid biosynthetic genes. J Biol Chem 267, 5751–5754.
    [Google Scholar]
  30. Reuhs, B. L., Geller, D. P., Kim, J. S., Fox, J. E., Kolli, V. S. & Pueppke, S. G. ( 1998; ). Sinorhizobium fredii and Sinorhizobium meliloti produce structurally conserved lipopolysaccharides and strain-specific K antigens. Appl Environ Microbiol 64, 4930–4938.
    [Google Scholar]
  31. Robertsen, B. K., Aman, P., Darvill, A. G., McNeil, M. & Albersheim, P. ( 1981; ). Host–symbiont interactions. V. The structure of acidic extracellular polysaccharides secreted by Rhizobium leguminosarum and Rhizobium trifolii. Plant Physiol 67, 389–400.[CrossRef]
    [Google Scholar]
  32. Rock, C. O. ( 2008; ). Fatty acids and phospholipids metabolism in prokaryotes. In Biochemistry of Lipids, Lipoproteins and Membranes, 5th edn, pp 59–96. Edited by D. E. Vance & J. E. Vance. Amsterdam: Elsevier.
  33. Sambrook, J. & Russell, D. R. ( 2001; ). Molecular Cloning: a Laboratory Manual. Cold Spring Harbor Laboratory, NY: Cold Spring Harbor Laboratory.
  34. Schäfer, A., Tauch, A., Jäger, W., Kalinowski, J., Thierbach, G. & Pühler, A. ( 1994; ). Small mobilizable multi-purpose cloning vectors derived from the Escherichia coli plasmids pK18 and pK19: selection of defined deletions in the chromosome of Corynebacterium glutamicum. Gene 145, 69–73.[CrossRef]
    [Google Scholar]
  35. Schägger, H. & von Jagow, G. ( 1987; ). Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem 166, 368–379.[CrossRef]
    [Google Scholar]
  36. Schweizer, H. D. ( 1993; ). Small broad-host-range gentamicin resistance gene cassettes for site-specific insertion and deletion mutagenesis. Biotechniques 15, 831–834.
    [Google Scholar]
  37. Sharypova, L. A., Niehaus, K., Scheidle, H., Holst, O. & Becker, A. ( 2003; ). Sinorhizobium meliloti acpXL mutant lacks the C28 hydroxylated fatty acid moiety of lipid A and does not express a slow migrating form of lipopolysaccharide. J Biol Chem 278, 12946–12954.[CrossRef]
    [Google Scholar]
  38. Sherwood, M. T. ( 1970; ). Improved synthetic medium for the growth of Rhizobium. J Appl Bacteriol 33, 708–713.[CrossRef]
    [Google Scholar]
  39. Simon, R., Priefer, U. & Pühler, A. ( 1983; ). A broad host range mobilization system for in vivo genetic engineering: transposon mutagenesis in Gram negative bacteria. BioTechnology 1, 784–791.[CrossRef]
    [Google Scholar]
  40. Sohlenkamp, C., de Rudder, K. E. & Geiger, O. ( 2004; ). Phosphatidylethanolamine is not essential for growth of Sinorhizobium meliloti on complex culture media. J Bacteriol 186, 1667–1677.[CrossRef]
    [Google Scholar]
  41. Studier, F. W., Rosenberg, A. H., Dunn, J. J. & Dubendorff, J. W. ( 1990; ). Use of a T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol 185, 60–89.
    [Google Scholar]
  42. Sun, Y., Hong, H., Gillies, F., Spencer, J. B. & Leadlay, P. F. ( 2008; ). Glyceryl-S-acyl carrier protein as an intermediate in the biosynthesis of tetronate antibiotics. ChemBioChem 9, 150–156.[CrossRef]
    [Google Scholar]
  43. Wu, K., Chung, L., Revill, W. P., Katz, L. & Reeves, C. D. ( 2000; ). The FK520 gene cluster of Streptomyces hygroscopicus var. ascomyceticus (ATCC 14891) contains genes for biosynthesis of unusual polyketide extender units. Gene 251, 81–90.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.033480-0
Loading
/content/journal/micro/10.1099/mic.0.033480-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error